LOCUS       Exported                7528 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZC320, complete sequence.
ACCESSION   U26464
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7528)
  AUTHORS   Shi J, Biek DP.
  TITLE     A versatile low-copy-number cloning vector derived from plasmid F
  JOURNAL   Gene 164 (1), 55-58 (1995)
  PUBMED    7590321
REFERENCE   2  (bases 1 to 7528)
  AUTHORS   Mori H, Kondo A, Ohshima A, Ogura T, Hiraga S.
  TITLE     Structure and function of the F plasmid genes essential for 
            partitioning
  JOURNAL   J. Mol. Biol. 192 (1), 1-15 (1986)
  PUBMED    3029390
REFERENCE   3  (bases 1 to 7528)
  AUTHORS   Prentki P, Binda A, Epstein A.
  TITLE     Plasmid vectors for selecting IS1-promoted deletions in cloned DNA: 
            sequence analysis of the omega interposon
  JOURNAL   Gene 103 (1), 17-23 (1991)
  PUBMED    1652541
REFERENCE   4  (bases 1 to 7528)
  AUTHORS   Biek DP.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-1995) Donald P. Biek, Department of Microbiology &
            Immunology, University of Kentucky Medical Center, 800 Rose Street, 
            Lexington, KY 40536-0084, USA
REFERENCE   5  (bases 1 to 7528)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7528
                     /organism="Cloning vector pZC320"
                     /plasmid="pZC320"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:40299"
     rep_origin      74..293
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             384..1139
                     /codon_start=1
                     /gene="repE"
                     /product="RepE"
                     /function="replication initiator protein"
                     /label=repE
                     /experiment="experimental evidence, no additional details 
                     recorded"
                     /protein_id="AAC53635.1"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     gene            384..1139
                     /gene="repE"
                     /label=repE
     misc_feature    1142..1392
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     repeat_region   1142..1389
                     /standard_name="incC"
                     /function="copy number control"
                     /label=incC
                     /rpt_family="iteron, RepE binding"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     CDS             1718..2893
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             1727..2893
                     /codon_start=1
                     /gene="sopA"
                     /product="SopA"
                     /function="plasmid partition"
                     /label=sopA
                     /experiment="experimental evidence, no additional details 
                     recorded"
                     /protein_id="AAC53636.1"
                     /translation="MKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRIGEA
                     ADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRAEDV
                     FPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDLHIH
                     AEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTDPHL
                     MLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFFDML
                     RDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDEVGK
                     GQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     gene            1727..2893
                     /gene="sopA"
                     /label=sopA
     CDS             2893..3864
                     /codon_start=1
                     /gene="sopB"
                     /product="SopB"
                     /function="plasmid partition"
                     /label=sopB
                     /experiment="experimental evidence, no additional details 
                     recorded"
                     /protein_id="AAC53637.1"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     gene            2893..3864
                     /gene="sopB"
                     /label=sopB
     repeat_region   3937..4454
                     /standard_name="sopC"
                     /function="plasmid partition centromere element"
                     /label=sopC
                     /rpt_family="SopB binding"
                     /rpt_unit_range=3981 .. 4023
                     /experiment="experimental evidence, no additional details 
                     recorded"
     misc_feature    3937..4410
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     regulatory      4785..4922
                     /regulatory_class="terminator"
                     /standard_name="omega terminator"
                     /note="contains the T4 gene 32 terminator and a portion of 
                     the polylinker from the omega interposon"
     CDS             complement(5249..6109)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /note="encoded on the SspI-BspHI bla fragment from pBR322; 
                     expressed from the aadA promoter"
                     /experiment="experimental evidence, no additional details 
                     recorded"
                     /protein_id="AAC53638.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5249..6109)
                     /gene="bla"
                     /label=bla
     CDS             complement(6122..6181)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside 3-acetyltransferase"
                     /number=1
                     /label=aadA
                     /note="encodes the first 18 amino acids of AadA and one 
                     from the vector before stopping 12 bp from the beginning of
                     bla coding region"
                     /protein_id="AAC53639.1"
                     /translation="MREAVIAEVSTQLSEVVGD"
     gene            complement(6122..6181)
                     /gene="aadA"
                     /label=aadA
     CDS             complement(6769..7071)
                     /codon_start=1
                     /gene="lacZa"
                     /product="beta-galactosidase alpha"
                     /function="beta-galactosidase alpha donor"
                     /number=1
                     /label=lacZa
                     /note="contains a multiple cloning site incorporated into 
                     the 5 end of lacZ. Following 45 codons in the MCS are lacZ
                     codons no.7-59 and then 2 codons from the vector before a 
                     UAA stop"
                     /experiment="experimental evidence, no additional details 
                     recorded"
                     /protein_id="AAC53640.1"
                     /translation="MTMITPSARNTTHYRAAASMLESSADVVRREAFERVCIDHVDPAR
                     SLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL"
     gene            complement(6769..7071)
                     /gene="lacZa"
                     /label=lacZa
     primer_bind     6918..6934
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7027..7045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7067..7083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7089..7201)
                     /standard_name="lacO1-lacO3"
                     /function="control of lacZa expression"
                     /label=lacO1-lacO3 binding site
                     /bound_moiety="lac repressor"
                     /note="The region upstream of lacZa contains the wild-type 
                     lac operator/promoter region from plasmid pBluescript SKII 
                     (Stratagene)"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     protein_bind    7091..7107
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7115..7145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7160..7181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      complement(7399..7528)
                     /regulatory_class="terminator"
                     /standard_name="omega terminator"
                     /note="contains the T4 gene 32 terminator and portion of 
                     the polylinker from the omega interposon"